Dako Autostainer - Manual (Page 108)

108 Dako Autostainer | User Guide
Phosphate Buffered Saline
The following is a procedure for the formulation of 20mM Phosphate, 150mM NaCl, buffered saline,
pH 7.0 with 0.05% Tween 20 from common laboratory chemicals.
Intended Use
A phosphate buffered saline solution, pH 7.0 can be used on the Dako Autostainer.
Materials Required
1. Clean carboy, flask or beaker graduated to 1L
2. Deionized water
3. Magnetic stir plate with stir bar
Common laboratory chemicals
Chemical Manufacturer and Product No.
Sodium Phosphate, dibasic (anhydrous) Sigma, S-087
Sodium Phosphate, monobasic (monohydrate) Fisher, S369-1
NaCl Sigma, S-9625
Tween 20 Dako, S1966
Sodium Hydroxide (NaOH), 50% w/w solution Baker, 3727-03
Reagent Preparation
One Liter of Phosphate-Buffered Saline containing 0.05% Tween 20
1. Add approximately 500 mL of deionized water to a 1L (1000 mL) carboy, flask or beaker.
2. Add 0.92 g of Sodium Phosphate, monobasic (monohydrate), 1.92 g of Sodium
Phosphate, dibasic and 8.77 g of NaCl and dissolve using a magnetic stir plate.
3. After all granules are dissolved adjust the solution to pH 7.0 with NaOH (50% w/w
solution). Allow the solution to continue to mix on the stir plate. Add the NaOH dropwise,
observing the pH constantly.
4. Once pH 7.0 is reached, add 0.5 mL of Tween 20 and mix using a magnetic stir plate.
5. Adjust the final volume of buffer to 1 liter with deionized water.
6. The final solution is 20mM Phosphate, 150mM NaCl, buffered saline, pH 7.0 with 0.05%
Tween 20.